The project seeks to explore the potential relationship of the non cAMP-dependent protein kinases in the DNA-dependent RNA polymerases I, II and III which transcribe RNA in the nucleus of the cell. To this end, the nuclear protein kinases are being purified to begin to study the types of proteins which are endogenous substrates for these enzymes. In addition, nuclear phosphatases are being characterized for their possible future. Experiments are also directed to the possible modulation of these nuclear protein kinases under conditions where changes in RNA polymerase activities are known to occur (e.g. hormones in target tissues). To date, nuclear protein kinase NII (eluted from DEAE columns) has been purified to homogeneity and nuclear protein kinase NI (found in DEAE flow-through) has been purified to about 2,000 fold. The properties of these enzymes are also being studied to compare the kinetic parameters of these enzymes to the cytoplasmic enzyme(s) which are cAMP dependent. It is hoped that these comparisons and properties will allow an assessment of whether there is a cytoplasmic-nuclear shuttle of protein kinase(s) (i.e., translocation). BIBLIOGRAPHIC REFERENCES: Zerwekh, J.E., Lindell, T.J. and Haussler, M.R., "Increased Intestinal Chromatin Template Activity: Influence of 1 alpha, 25-Dihydroxyvitamin D3 and Hormone Receptor Complexes," J. Biol. Chem. 251, 2388 (1976). Lindell, T.J., Ellinger, R., Warren, J.F., Sundheimer, D., and O'Malley, A.F., "The Effect of Acute and Chronic Phenobarbital Treatment on the Activity of Rat Liver DNA-Dependent RNA Polymerases," Molecular Pharmacology, May 1977, in press.